百草枯诱导Ⅱ型肺泡上皮细胞TGF-β1的变化及甘草酸二铵干预作用  

Effects of Diammonium Glycyrrhizinate on the Expression of TGF-β1 in Alveolar Epithelial Cell Ⅱ cultured with Paraquat

作　　者：向沥 方辰[1] 周志兵 张剑锋[1] XIANG Li;FANG Chen;ZHOU Zhi-bing;ZHANG Jian-feng(Department of General medicine,Second Affiliated Hospital of Guangxi Medical University,Nanning,Guangxi,530007,China)

机构地区：[1]广西医科大学第二附属医院全科医学科,广西南宁530007

出　　处：《蛇志》2018年第1期7-9,17,共4页Journal of Snake

基　　金：广西重点研发计划项目(桂科AB17195002);广西自然科学基金项目(2017GXNSFAA198249);广西医疗卫生适宜技术开发与推广应用项目(S2017009);广西高校本科教学改革工程项目(02506217091C)

摘　　要：目的探讨甘草酸二铵(DG)对百草枯(PQ)刺激大鼠Ⅱ型肺泡上皮细胞(alveolar epithelial cellⅡ,AECⅡ)分泌TGF-β1的影响及其机制。方法采用不同浓度的PQ溶液(200、400、600、800、1000、1500、2000μmol/L)诱导大鼠Ⅱ型肺泡上皮细胞24h,测定PQ对AECⅡ生存率的影响,了解IC50的PQ浓度。在1000μmol/L PQ诱导下,以0、0.2、0.4、0.6、0.8、1.0、2.0mg/ml浓度DG干预AECⅡ24h,测定DG对PQ诱导AECⅡ生存率的影响,了解DG干预PQ诱导下AECⅡ最高生存率的DG浓度。然后将AECⅡ分为3组,即空白对照组、PQ组(以1000μmol/L PQ培养24h)、DG组(先以0.6mg/ml DG培养2h,再以0.6mg/ml DG+1000μmol/L PQ培养24h),使用倒置显微镜观察AECⅡ的形态改变、生长情况,采用酶联免疫吸附法检测TGF-β1的含量,采用实时荧光定量聚合酶链式反应测定TGF-β1mRNA的表达。结果 IC50的PQ浓度为920.87μmol/L,在1000μmol/L PQ诱导下,以0.6mg/ml浓度DG干预AECⅡ24h的生存率最高。与空白对照组比较,PQ组TGF-β1的含量及其mRNA的表达水平明显升高,差异有统计学意义(P<0.05);与PQ组比较,DG组TGF-β1的含量及其mRNA的表达水平明显下降,差异有统计学意义(P<0.05)。结论本实验成功建立了合适的PQ损伤大鼠Ⅱ型肺泡上皮细胞的模型及DG干预模型。甘草酸二铵可以降低百草枯诱导下的AECⅡ分泌TGF-β1水平。Objective To investigate the effects of Diammonium Glycyrrhizinate(DG)on the expression of TGF-β1 in rat alveolar epithelial cellⅡ(AECⅡ)cultured with paraquat(PQ). Methods Rat alveolar epithelial cellⅡwere cultured with varied concentrations(200,400,600,800,1000,1500,2000μmol/L)of PQ for 24 h.The viability of cells was measured to know the concentrations of PQ at IC 50.Rat AECⅡcultured with 1000μmol/L PQ were treated with varied concentrations(0,0.2,0.4,0.6,0.8,1.0,2.0 mg/ml)of DG for 24 h.The viability of cells treated with DG was measured to know the suitable concentrations of DG.The cells were divided into three groups:The NS group was cultured 24 h without any drugs,the PQ group was cultured 24 h with 1000μmol/L PQ,and the DG group was treated with 0.6 mg/ml DG for 2 h,then cultured with 0.6mg/ml DG+1000μmol/L PQ for 24 h.Growth status of AECⅡwas observed by inverted microscope;the ELISA assay was applied to measure the levels of TGF-β1 in the three groups.The gene expressions of TGF-β1 mRNA in the three groups were detected by RT-PCR. Results The concentrations of PQ at IC 50 was 920.87μmol/L.The viability of AECⅡcultured with 1000μmol/L PQ was the hightest when treated with 0.6 mg/ml of DG;The levels of TGF-β1 in PQ group were higher than those in NS group.While,the levels of TGF-β1 in DG group were lower than those in PQ group(P<0.05).The expression of TGF-β1 mRNA in PQ group were higher than those in NS group.The expression of TGF-β1 mRNA in DG group were lower than those in PQ group(P<0.05). Conclusion An appropriate vitro model of AECⅡcultured with PQ and treated with DG have been successfully established in present study.We found DG can decrease the expressions of TGF-β1 in AECⅡcultured with PQ.

关 键 词：甘草酸二铵 百草枯 Ⅱ型肺泡上皮细胞 转化生长因子Β1 

分 类 号：R341.31[医药卫生—基础医学]